Optogenetics: Background, Methodological Advances and Potential Applications for Cardiovascular Research and Medicine

Optogenetics is an elegant approach of precisely controlling and monitoring the biological functions of a cell, group of cells, tissues, or organs with high temporal and spatial resolution by using optical system and genetic engineering technologies. The field evolved with the need to precisely control neurons and decipher neural circuity and has made great accomplishments in neuroscience. It also evolved in cardiovascular research almost a decade ago and has made considerable progress in both in vitro and in vivo animal studies. Thus, this review is written with an objective to provide information on the evolution, background, methodical advances, and potential scope of the field for cardiovascular research and medicine. We begin with a review of literatures on optogenetic proteins related to their origin, structure, types, mechanism of action, methods to improve their performance, and the delivery vehicles and methods to express such proteins on target cells and tissues for cardiovascular research. Next, we reviewed historical and recent literatures to demonstrate the scope of optogenetics for cardiovascular research and regenerative medicine and examined that cardiac optogenetics is vital in mimicking heart diseases, understanding the mechanisms of disease progression and also in introducing novel therapies to treat cardiac abnormalities, such as arrhythmias. We also reviewed optogenetics as promising tools in providing high-throughput data for cardiotoxicity screening in drug development and also in deciphering dynamic roles of signaling moieties in cell signaling. Finally, we put forth considerations on the need of scaling up of the optogenetic system, clinically relevant in vivo and in silico models, light attenuation issues, and concerns over the level, immune reactions, toxicity, and ectopic expression with opsin expression. Detailed investigations on such considerations would accelerate the translation of cardiac optogenetics from present in vitro and in vivo animal studies to clinical therapies

Impulse oscillometry

Pulmonary function testing involves a battery of tests from the simple pulse oximetry to the cardiopulmonary exercise testing. Impulse oscillometry (IOS) is one of the newly described pulmonary function tests. It is based on the old principle of forced oscillatory technique modified and refined as per research and advances. It involves the use of sound waves during normal tidal breathing, which gives information on oscillatory pressure-flow relationships and eventually resistance and reactance. The resistance at 20 Hz (R20) represents the resistance of the large airways. The resistance at 5 Hz (R5) means the total airway resistance. (R5–R20) reflects resistance in the small airways. The reactance at 5 Hz (X5) indicates the elastic recoil of the peripheral airways. Resonant frequency and area of reactance are also measured. IOS has major uses in diagnosis and control of asthma in children and the elderly, where spirometry is otherwise normal. IOS has been studied in other respiratory diseases like COPD, ILD and supraglottic stenosis

Synthesis and Secretome Release by Human Bone Marrow Mesenchymal Stem Cell Spheroids within Three-dimensional Collagen Hydrogels: Integrating Experiments and Modelling

Myocardial infarction results in loss of cardiac cell types, inflammation, extracellular matrix (ECM) degradation, and fibrotic scar. Transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) is being explored as they could differentiate into cardiomyocyte-like cells, integrate into host tissue, and enhance resident cell activity. The ability of these cells to restore lost ECM, remodel the inflammatory scar tissue, and repair the injured myocardium remains unexplored. We here elucidated the synthesis and deposition of ECM (e.g., elastin, sulfated glycosaminoglycans, hyaluronan, collagen type III, laminin, fibrillin, lysyl oxidase, and nitric oxide synthases), matrix metalloproteinases (MMPs) and their inhibitors (TIMPs), and other secretome (cytokines, chemokines, and growth factors) in adult human BM-MSC spheroid cultures within three-dimensional collagen gels. The roles of species-specific type I collagen and 5-azacytadine were assessed over a 28-day period. Results revealed that human collagen (but not rat-derived) suppressed MSC proliferation and survival, and MSCs synthesized and released a variety of ECM proteins and secretome over the 28 days. Matrix deposition is at least an order of magnitude lower than their release levels at every time point, most possibly due to elevated MMP levels and interleukins with a concomitant decrease in TIMPs. Matrix synthesis over the 28-day period was fitted to a competitive inhibition form of Michaelis-Menten kinetics, and the production and decay rates of ECM, MMPs, and TIMPs, along with the kinetic model parameters quantified. Such an integrated experimental and modelling approach would help elucidate the critical roles of various parameters (e.g., cell encapsulation and delivery vehicles) in stem cell-based transplantation therapies

Role of GeneXpert in the diagnosis of mycobacterium tuberculosis

Introduction: GeneXpert (GX) is a novel, integrated, cartridge-based, nucleic acid amplification test with an established role for rapid diagnosis of Mycobacterium tuberculosis and detection of rifampicin resistance.Aim: To evaluate the role of GX in pulmonary and extrapulmonary tuberculosis (TB) cases.Material and methods: A prospective study was conducted in the pulmonary medicine department of a tertiary care hospital after the Ethics Comittee permission. Data of 257 presumptive TB patients was retrieved for GX, acid fast bacilli smear and cul-ture (AFB smear and culture) and drug susceptibility test (DST). Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of GX in diagnosis and determination of rifampicin resistance in pulmonary and extrapulmonary TB cases were calculated and compared with culture and DST results.Results: Our study included 132 pulmonary and 125 extrapulmonary cases. On the basis of clinicoradiological and microbiological correlation, diagnosis of TB was confirmed in 104 pulmonary and 103 extrapulmonary cases. Out of a total of 104 pulmonary TB cases, 73 were rifampicin-sensitive and 31 were rifampicin-resistant cases. 103 extrapulmonary TB patients included 66 rifampicin-sensitive and 37 rifampicin-resistant cases. The sensitivity, specificity, PPV, NPV of GX in diagnosis and detection of rifampicin resistance in pulmonary TB was 95%, 93%, 98%, 84% and 96%, 100%, 100%, 96%, respectively. The sensitivity, specificity, PPV, NPV of GX in diagnosis and detection of rifampicin resistance in extrapulmonary TB cases was 79%, 86%, 96%, 47% and 97%, 95%, 97%, 95%, respectively.Conclusions: GX results are superior to smear microscopy and comparable to culture with shorter turnaround time.We recom-mend using it in routine TB diagnosis as this will expedite the management of patients with presumptive TB

Fe(III)-Chloroquine Complex: A New Potent Compound in Wellness Industries of High Antimicrobial and Antimalarial Activities

Fe(III)- Chloroquine complex has been synthesized and screened for its physicochemical, microbial as well as pharmacological activity have been done in solid and aqueous phase. On the basis of elemental analysis, polarographic studies, amperometric titration and IR spectral studies the probable formula for the complex has been determined at 25±1OC and ionic strength of µ= 1.0[KCl]. Raper's paper disc method was used for microbial study against various pathogenic bacteria and fungi. In vivo syudy of Swiss mice [25-30gm] were used for antimalarial activity against Chloroquine and its complex on xyline-Alcoholic activity test Kidney, liver and serum of these rats were also studied. On the basis of observed result it could be concluded that Fe (III)-Chloroquine complex were found to be non-toxic and more potent than pure chloroquine drug

Role of Metals in Pediatric Oral Health

Prefabricated stainless steel crowns (SSCs) are the regular dental prosthesis cemented to primary molars in children. Previously used SSC, which contained up to 72% nickel, is associated with nickel sensitivity. Hence, the new generation of SSC that contains only 9–12% nickel was developed. Stainless steel orthodontic materials and stainless steel crowns (SSC) are the two major devices in pediatric patients that contain heavy metals. Measurable amounts of nickel and chromium in the saliva and serum are released from this prosthesis without reaching toxic levels. Allergic reaction in a form of gingivitis was reported after 3 months in 20% of the females and 10% of the males, and it disappeared a month after appliance removal. Several studies reported that there is more leaching of metals in acidic pH. Many different types of alloys are now available in the market to be used for dental restorations and fixed prostheses, and the rates of metal leaching from these alloys are not known. The common criterion for all these fixed prosthodontic materials is their permanent existence in the oral cavity for a prolonged time without the ability to be removed by the patient. Let us know these elements in detail in this chapter

Alterations in phenotype and gene expression of adult human aneurysmal smooth muscle cells by exogenous nitric oxide

International audienceAbdominal aortic aneurysms (AAA) are characterized by matrix remodeling, elastin degradation, absence of nitric oxide (NO) signaling, and inflammation, influencing smooth muscle cell (SMC) phenotype and gene expression. Little is known about the biomolecular release and intrinsic biomechanics of human AAA-SMCs. NO delivery could be an attractive therapeutic strategy to restore lost functionality of AAA-SMCs by inhibiting inflammation and cell stiffening. We aim to establish the differences in phenotype and gene expression of adult human AAA-SMCs from healthy SMCs. Based on our previous study which showed benefits of optimal NO dosage delivered via S-Nitrosoglutathione (GSNO) to healthy aortic SMCs, we tested whether such benefits would occur in AAA-SMCs. The mRNA expression of three genes involved in matrix degradation (ACE, ADAMTS5 and ADAMTS8) was significantly downregulated in AAA-SMCs. Total protein and glycosaminoglycans synthesis were higher in AAA-SMCs than healthy-SMCs (p < 0.05 for AAA-vs. healthy-SMC cultures) and was enhanced by GSNO and 3D cultures (p < 0.05 for 3D vs. 2D cultures; p < 0.05 for GSNO vs. non-GSNO cases). Elastin gene expression, synthesis and deposition, desmosine crosslinker levels, and lysyl oxidase (LOX) functional activity were lower, while cell proliferation, iNOS, LOX and fibrillin-1 gene expressions were higher in AAA-SMCs (p < 0.05 between respective cases), with differential benefits from GSNO exposure. GSNO and 3D cultures reduced MMPs −2, −9, and increased TIMP-1 release in AAA-SMC cultures (p < 0.05 for GSNO vs. non-GSNO cultures). AAA-SMCs were inherently stiffer and had smoother surface than healthy SMCs (p < 0.01 in both cases), but GSNO reduced stiffness (~25%; p < 0.01) and increased roughness (p < 0.05) of both cell types. In conclusion, exogenously-delivered NO offers an attractive strategy by providing therapeutic benefits to AAA-SMCs

Diffuse Pan Bronchiolitis Presenting with Bronchiectasis: A Case Report

Diffuse Pan Bronchiolitis (DPB) is a peculiar airway disease with its pathogenesis enrooted in a complex interplay of various genetic and environmental factors. Airway inflammation, chronic airflow limitation and suppuratives in pulmonary infections are the distinctive features of this entity. It poses a close differential to other frequently encountered pulmonary conditions like chronic bronchitis, emphysema, bronchiectasis and constrictive bronchiolitis. Deferment in diagnosis can culminate in irreversible airway remodeling and progressive respiratory failure. Hence a punctual recognition is vital. Macrolide group of drugs are prime modality of therapy and the response to therapy is benignant. We herein describe a case of DPB with development of sequelae owing to its delayed detection

Suffering in silence: consequences of sexual violence within marriage among young women in Nepal

<p>Abstract</p> <p>Background</p> <p>Despite the grave consequences of sexual violence, and it's persistence both within and outside marriages, this subject has received relatively little attention from researchers, policy makers, and programme managers in Nepal. This paper explores the definition of sexual violence and its various forms and consequences as reported by young married women in Nepal. In addition, it describes the coping mechanisms used by young married women to avoid sexual violence perpetrated against them by their husbands.</p> <p>Methods</p> <p>This paper analyzes data collected during the qualitative study on "Sexual violence among young couples in Nepal", conducted amongst two major ethnic groups - Brahmin/Chhetri and Tharu - between 2006 and 2007. The data is comprised of 39 free-lists and 15 in-depth case histories with married women aged 15-24 years. The average rank and Smith's Salience were calculated from the free-listing data. The thematic analysis approach was used for the analysis of information from the case histories.</p> <p>Results</p> <p>Approximately three-quarters (74%) of the young women mentioned 'sex against one's wishes' as sexual violence within marriage (SVWM). Sixty-two percent of respondents described 'forced sex during illness' and 'forced sex after consumption of alcohol' as SVWM. About half of young women (48.7%) who participated in the free-listing exercise reported having experienced SVWM. The types of SVWM ranged from unwanted sexual touch to forced sex. Backache, headache, lower abdominal pain, vaginal bleeding and thoughts of suicide were the most commonly reported negative physical and psychological health consequences of SVWM. Women reported various coping strategies including 'trying to convince husbands', 'sleeping in a separate room', 'visiting maternal home', 'waking up the children', and 'using pretexts such as being ill or menstruating', to avoid SVWM. However, in most cases, women reported that these coping strategies were unsuccessful. Almost all women experiencing SVWM were socially isolated and did not turn to institutions, relatives or friends for advice and support.</p> <p>Conclusions</p> <p>Sexual violence within marriage is common in Nepal. Findings provide circumstantial evidence of links between sexual violence and negative general and reproductive health outcomes for women. Various actions are required to prevent SVWM and provide immediate support to the victims.</p